Biofilms possess unique mechanical properties that are vital to their stability and function. Biofilms are made of extracellular polymeric substances (EPS) secreted by microorganisms and comprise polysaccharides, proteins, extracellular DNA (eDNA), and lipids. EPS is the primary contributor and driver of the biofilm structure and mechanical properties such as stiffness, cohesion, and adhesion. EPS enhances the elasticity and viscosity of biofilms, allowing them to withstand mechanical stresses, shear forces, and deformation. Therefore, biofilms are notoriously difficult to remove and can result in billions of dollars in losses for various industries due to their adverse effects, such as contamination, pressure loss, and corrosion. As a result, it is essential to comprehend the mechanical properties of biofilms to control or remove them in various scenarios. We undertook a fundamental study to determine the relationship between individual EPS components and biofilm mechanical properties. In this study, a CDC biofilm reactor was used to grow pure culture biofilms (Staphylococcus epidermidis) which were treated with six EPS modifier agents (Ca²⁺, Mg²⁺, periodic acid, protease K, lipase, and DNAase I) to modify or cleave specific EPS components. The mechanical properties (Young's Modulus) of treated biofilms were subsequently tested using atomic force microscopy (AFM), the biofilm EPS functional groups were measured via the Fourier transform infrared (FTIR) spectroscopy, and biofilm structural characteristics using confocal imaging. The FTIR results showed that EPS modifier agents successfully reduced their target EPS components. Similarly, the confocal microscopic analysis results showed that most of these modifier agents (except lipase) significantly reduced (P-value <0.05) the biovolume and thickness of treated biofilms. Conversely, most of these modifier agents (except protease K) significantly increased (P-value <0.05) the roughness coefficient of the biofilms. Finally, data from AFM showed that biofilm mechanical properties (Young's modulus) significantly (P-value <0.05) changed with their EPS composition. These results have significant ramifications for biofilm management and control in myriad scenarios.

01 Oct 2024·International Journal of Biological Macromolecules

Lipase-copper complex/chitosan microspheres; loaded with attapulgite used for the treatment of E. coli-induced diarrhea

Article

Author: Dureshahwar ; Khan, Bushra ; Ameer, Nabeela ; Arif, Aimen ; Shah, Amir Asad ; Hanif, Muhammad ; Khan, Waheed S ; Mahmood, Khalid ; Arif, Maheen ; Nisar, Hafiza Razia

Diarrhea is a globally major problem especially Escherichia coli induced diarrhea becoming fatal nowadays in developing countries. Colon-targeted chitosan microspheres (Ms) comprising of lipase‑zinc and lipase‑copper complexes were prepared, loaded with Attapulgite (Cts-Li-Zn-ATG/Ms and Cts-Li-Cu-ATG/Ms) for the treatment of bacterial diarrhea. Thin layer chromatography (TLC) and Fourier-transform infrared spectroscopy (FTIR) studies were used for confirmation of proposed lipase-metal complexes. Ms showed particle size range 18 ± 0.24 to 23 ± 0.83 μm, zeta potential -13.7 ± 0.71 to -29.3 ± 1.34 mV, PDI 0.5 ± 0.04 to 1.0 ± 0.07 and hemolytic activity was found to be <5 ± 1.25 %. After coating with Eudragit S-100 for colon targeting, in-vitro % drug release of ATG at pH 7.4 was 80 ± 0.21 % for Eud-Cts-Li-Zn-ATG/Ms while it was increased to 83 ± 0.54 % for Eud-Cts-Li-Cu-ATG/Ms within 7 h, respectively. In-vivo anti-diarrheal activity of Eud-Cts-Li-Zn-ATG/Ms and Eud-Cts-Li-Cu-ATG/Ms was performed by oral challenge on albino mice having infectious diarrhea colonized with E. coli. Results revealed significant anti-diarrheal effect of proposed Eud-Cts-Li-Cu-ATG/Ms in terms of weight gain from 24 ± 0.12 g to 26.05 ± 0.31 g, which was 2-fold increase as compared to Eud-Cts-Li-Zn-ATG/Ms. Conclusively, Eud-Cts-Li-Cu-ATG/Ms provides an innovative alternate for the treatment of bacterial diarrhea with additional support of chitosan and lipase for nutritional support and immunity which was compromised in diarrheal patients.

25 Sep 2024·Clinical Chemistry and Laboratory Medicine (CCLM)

Sharing reference intervals and monitoring patients across laboratories – findings from a likely commutable external quality assurance program

Article

Author: Farrell, Christopher J.L. ; Jones, Graham R.D. ; Badrick, Tony ; Sikaris, Kenneth A. ; Graham, Peter ; Bush, Jonathan

AbstractObjectivesLaboratory results are increasingly interpreted against common reference intervals (CRIs), published clinical decision limits, or previous results for the same patient performed at different laboratories. However, there are no established systems to determine whether current analytical performance justifies these interpretations. We analysed data from a likely commutable external quality assurance program (EQA) to assess these interpretations.MethodsThe use of CRIs was assessed by evaluating instrument group medians against minimum specifications for bias. The use of clinical decision limits was assessed using specifications from professional bodies, and the monitoring of patients by testing at different laboratories was assessed by comparing all-laboratory imprecision to within-subject biological variation.ResultsFive of the 18 analytes with Australasian CRIs did not meet specification for all instrument groups. Among these, calcium and magnesium failed for one instrument group out of seven, while bicarbonate, chloride, and lipase failed for two instrument groups. Of the 18 analytes reviewed currently without CRIs in Australasia, 10 candidates were identified. Among analytes with clinical decision limits, i.e. lipids, glucose, and vitamin D, only triglycerides met both bias and imprecision specifications, while vitamin D met the imprecision specification. Monitoring patients by testing at different laboratories was supported for 15 of the 46 (33 %) analyte-method principles groups that met minimum imprecision specifications.ConclusionsAnalysis of data from commutable EQA programs can provide a mechanism for monitoring whether analytical performance justifies the interpretations made in contemporary laboratory practice. EQA providers should establish systems for routinely providing this information to the laboratory community.

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